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. Author manuscript; available in PMC: 2021 Nov 1.
Published in final edited form as: Biochim Biophys Acta Mol Basis Dis. 2020 Jul 22;1866(11):165901. doi: 10.1016/j.bbadis.2020.165901

Figure 1. Effect of FICZ on Th17 effector cytokines IL-17 and IL-22 release in T cells after ethanol and burn injury.

Figure 1.

One day after injury, mice were euthanized and distal small intestine was collected for LP cell isolation. Isolated LP cells (1x106) were stimulated with PMA (10ng/ml) and Ionomycin (50ng/ml) in cell culture medium for 2 h. GolgiPlug Protein Transport inhibitor (1 μl/ml) was add in culture medium for another 3 h. The cells were harvested and stained with Fixable Viability Dye, APC-eFluor780 Anti-mouse CD3, eFluor450 anti-mouse-IL-17 and PE anti-mouse IL-22 antibodies. LP cells were gated for live/CD3+ cells and then gated for IL-17 and IL-22 (Fig. 1A). The percentage of IL-22+ cells is shown in Fig. 1B and IL-17+ cells is shown in Fig. 1C. Spleen T (5 x 105 cells/well) were cultured with plate-bound anti-CD3 (5μg/ml) and anti-CD28 (1μg/ml) for 48 h and supernatants were collected to determine IL-22 (Fig. 1D), IL-17 (Fig. 1E), and CYP1A1 mRNA expression (Fig. 1F). Values are means ± SEM from four to eight animals per group (In Fig. 1A, B and C, sham vehicle n = 5, sham vehicle + FICZ n = 5, burn ethanol n = 8 and burn ethanol + FICZ n = 7. In Fig. 1D, E, and F, sham vehicle n = 5, sham vehicle + FICZ n = 4, burn ethanol n = 7 and burn ethanol + FICZ n = 6). P*<0.05 compared to other groups.